关键词: beans entomopathogenic fungus group I introns molecular detection nested PCR

来  源:   DOI:10.3390/plants13010022   PDF(Pubmed)

Abstract:
Beauveria bassiana (B. bassiana) is a significant entomopathogenic fungus (EPF) in agriculture as a sprayable biocontrol agent. It has the potential to be established as an endophyte (ENP) in various crops, resulting in beneficial effects for the host plants, including resistance to pest insects and increased growth and yield. However, it is not known whether a B. bassiana strain has such a favorable impact on the plant, since it is a common soil microorganism. Therefore, techniques that allow strain monitoring will be advantageous. To date, methods for detecting or monitoring a specific EPF strain after external application are scarce. In the present study, an in planta nested PCR technique was standardized to differentiate between three B. bassiana strains (GHA, PTG4, and BB37) established as endophytes in bean plants under laboratory conditions by detecting the insertion profile of four group I introns located in the 28S gene of B. bassiana ribosomal DNA. This technique recognized a distinct pattern of bands of different sizes for each strain, with a sensitivity of 1 pg per 10 ng of plant DNA. This molecular approach may be more effective monitoring B. bassiana strains after application to evaluate their significance on crops.
摘要:
白僵菌(B.bassiana)是农业中一种重要的昆虫病原真菌(EPF),可作为可喷雾的生物防治剂。它有可能在各种作物中被确立为内生菌(ENP),对寄主植物产生有益的影响,包括对害虫的抗性以及增加的生长和产量。然而,尚不清楚白孢芽孢杆菌菌株是否对植物具有如此有利的影响,因为它是一种常见的土壤微生物。因此,允许应变监测的技术将是有利的。迄今为止,在外部应用后检测或监测特定EPF菌株的方法很少。在本研究中,对植物巢式PCR技术进行了标准化,以区分三种玄武岩菌株(GHA,PTG4和BB37)在实验室条件下通过检测位于球孢芽孢杆菌核糖体DNA28S基因中的四个I组内含子的插入谱而在豆科植物中建立为内生菌。该技术识别出每种菌株不同大小的条带的独特模式,每10ng植物DNA的灵敏度为1pg。这种分子方法可能会更有效地监测施用后的白杨菌株,以评估其对作物的重要性。
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